不同体积分数血清对培养原代破骨细胞噬骨能力的影响  

Effect of serum of different volume fractions on resorption ability of primary osteoclasts

作　　者：柴立民[1] 王正东[2] 颜南[3] 

机构地区：[1]沈阳医学院高职学院教务科,辽宁省沈阳市110034 [2]沈阳医学院解剖教研室,辽宁省沈阳市110034 [3]沈阳医学院公共卫生学院学生科,辽宁省沈阳市110034

出　　处：《中国组织工程研究与临床康复》2007年第28期5485-5488,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:改变培养液中血清的浓度,观察破骨细胞形态结构,并比较不同体积分数血清对培养的原代破骨细胞噬骨能力的影响。方法:实验于2007-03/06在沈阳医学院中心实验室完成。①实验材料:选取清洁级新生24hWistar大鼠6只;胎牛血清(天津灏洋);新鲜成年牛股骨皮质(市售)。②实验过程:取新鲜成年牛股骨皮质,用磨片机磨成厚50μm的1cm×1cm骨片;取新生大鼠四肢长骨,分离破骨细胞,并用不同体积分数血清(分别为0,0.05,0.1,0.15,0.2,0.25)培养原代破骨细胞。③实验评估:第1,6天倒置显微镜下计数,第3天取细胞玻片观察细胞形态结构;扫描电镜观察不同体积分数血清培养的破骨细胞在牛骨皮质上产生骨陷凹面积的差异。结果:①破骨细胞的分离培养结果:分离培养的破骨细胞数量较少,体积大,胞浆丰满,细胞突起延展,细胞核呈圆形或椭圆形,积聚在细胞中央或排列在细胞周边。②不同体积分数血清培养液中骨片上吸收陷窝深度比较:随着培养液中血清体积分数的增加,骨片上陷窝深度逐渐增加,在培养液中血清体积分数为0.15时,骨片上陷窝最深,此后随着培养基血清体积分数的增加,形成骨陷凹的深度无明显增加。结论:血清体积分数为0.15培养基培养的破骨细胞在牛皮质骨片上产生骨陷凹最深,噬骨能力最强。AIM： To alter the serum concentration in culture solution, and then observe the osteoclasts structure and compare the effect of serum of different volume fractions on bone resorption of primary osteoclasts. METHODS： The experiment was performed at the Central Laboratory of Shenyang Medical College from March to June 2007. ①Six clean-grade Wistar rats aged 24 hours ware selected. Fetal calf serum （FCS） was bought from Tianjin Haoyang. Fresh adult calf femur cortex was purchased in market. ②Fresh adult calf femur cortex was grinded into 50μm thick 1 cm×1 cm specula with ground section machine. Long bone of four limbs was gotten from neonate rats, and then osteoclasts were isolated and cultured with serum of different volume fractions （0, 0.05, 0.1, 0.15, 0.2, 0.25）. ③ Osteoclasts were counted under inverted microscope at days 1 and 6. Cell slide was obtained at day 3 to observe cell appearance. The differences of bone excavation areas in calf bone cortex induced by osteoclasts cultured with serum of different volume fractions were observed by scanning electron microscope （SEM）. RESULTS： ①The osteoclasts ware not so much, and they were big in volume and with much kytoplasm, had cell process, the nucleus ware round or ellipse, and accumulated in the middle of cell or in the perimeter of the cell. ②With the increase of volume fraction of serum, the lacune deepened in specula. At the 0.15 volume fraction, the lacune was the deepest. When serum volume fraction was increased, the depth of resorption lacuna did not increase. CONCLUSION： The culture with serum of 0.15 volume fraction has the strongest ability of bone resorption, showing the lacune was the deepest.

关 键 词：巨细胞 破骨细胞 细胞培养技术 血清 组织构建 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]